Diastatic product and the process for producing the same.



" UNITED STATES PATENT orrron.

JOKIGHI TAKAMINE, OF NEW YORK, N. Y.

DIASTATIC PRODUCT AND THE PROCESS FOR PRODUCING THE SAME.

80 Drawing.

To all whom it may concern:

Be it known that I, JOKICHI TAKAMINE, a subject of the Emperor of Japan, and a resident of New York city, borough of Manhattan, county of New York, and State of New York, have invented a new and useful Diastatic Product and the Process for Producing the Same, of which the following is a s ecification n my Letters Patent of the United States numbered 525820, 525822 and 525823, I have described a diastase producing fungus and my present invention consists of an improvement thereon.

Heretofore, the products described in said patents have not gone into use commercially as malt substitutes in the brewing and alcoholic industries, because the products of said, patents have never heretofore possessed a diastatic strength superior to malt.

The product constituting my present invention possesses a diastatic strength when properly manufactured of from 30 to 50% in excess of malt. Therefore, my present invention constitutes a malt substitute as an article of manufacture capable of commercial use in competition with malt.

My product is produced by the growth under certain conditions of the spores of microscopic mycelial fungi which when sown on suitable material have the property of producing diastase. I employ the genus Aspergz'llus and particularly prefer the species Aspergillus org ace of pure culture because it excels any other species with regard to production of diastase in abundance. Therefore, in the following examples of my invention, I will employ for convenience,

Aspergz'llus org 2w, but do not wish to be understood as thereby limiting myself thereto.

Example of acalimatiaz'ng the spores.I take a suitable culture medium, such as wheat bran, moistened with water so as to contain from to 80 parts by wei ht of water to bran. This is sterilized an after cooling to about 30 C. an antiseptic, such as formaldehyde, is added in about the proportion of one part formaldehyde to 3000 arts of the moistened medium. A ure c ture of Aspergillus org 20c is intro uced upon the medium and the whole is kept in an incubator at a temperature of from 28 to 30 C. which latter is the optimum temperature for the growth of Aspergillus org 2w. When the growth is completed and the greenish yellow spores are produced a culture of the same Specification of Letters Patent.

Patented Aug. 3, 1915.

Application filed February 2, 1910. Serial No. 541,817.

is introduced upon a fresh culture medium of the same composition and containing the same proportion of formaldehyde.

At least four or five generations of the growth are produced in the same manner, employing substantially the same proportion of formaldehyde. In this manner, the spores become acclimated to that proportion of formaldehyde or substantially immune against its effects. Subsequent generations of the growth are then produced under an increase of the proportion of formaldehyde. Thus, a culture of the previously produced spores is introduced upon a fresh culture medium made as above excepting that the proportion of antiseptic is about one part of formaldehyde to 2900 parts of the moist medium. At least four or five generations of growth are produced at this proportion of antiseptic.

Similar operations are repeated until the proportion of antiseptic has increased to about one part of formaldehyde to 1500 parts of the moist medium each decrement in the proportion of moist medium being about 100 parts. By this gradual and step by step increase of the antiseptic, the spores become acclimatized or substantially immune to the proportion of antiseptic at each step before the next succeeding step is taken and in this way spores which, at the outset, would not have grown in the presence of a proportion of 1 to 1500 or less formaldehyde, attain the capacity of satisfactory growth under those conditions. When the maximum acclimatization has been reached it can be determined by observing that any further increase in the proportion of antiseptic pre- -vents the growth of the fungi. Having reached the maximum acclimatization when any further increase of proportion of antiseptic would interfere with the growth of the fungus, the spores contained in the prodarated spores being obtained as microscopic particles of a greenish yellow color..

Ewample of growth from cwclimatized spore8.The acclimatized spores obtained by the foregoing operations, either so arated from the culture medium on which they were produced or not so arated from the culture medium, are intro uced upon a new culture medium composed as before of sterilized and cooled wheat bran comly abundance of air;

bined with 60 to 80 parts by weight of Water and one part or less b weight of formaldehyde to 1500 piarts of t e moist medium, and the whole ept in an incubator at a temperature of from 28 to 30 C. (the optimum temperature for the growth of Aspergillus org aw) until the growth is complete. If the acclimatized spores beemployed, separated from the old culture medium, the proportion of them to the bran in the new culture medium exclusive of water, will be as 1 to 1500. If, however, the acclimatized spores be used in conjunction with the old culture medium, this proportion will be as 1 to 200. The whole is then dried at not to exceed 40 C. When completely dry it can be kept indefinitely. The dried mass may then be powdered and sold as an article of manufacture in that condition, or before sale the spores may be separated from the culture medium b sieving and the spores sold as pure. n either case, if produced in large quantities it constitutes the new malt substitute having from 30 to diastatic strength in excess of malt. It can be used in the brewing and alcoholic industries as a substitute for malt in accordance with another Patent No. 1,054,626, dated February 13, 1913, for making diastase of extraordinary strength in accordance with another application filed July 1, 1912, Serial No. 707,053.

Whether the resulting product be a new moyashi (Patent 525822) or a new koji (Patent 525820) depends upon the stage at which the final culture is interrupted. The intent is to cover both of them in the appended claims. At the commencement of the final culture, the temperature in the incubator will be at about 30 C. This gradually rises as the growth proceeds until it attains a temperature of 40 to 43 C. when the growth proceeds at maximum s eed. At t e end of about 48 hours, the cu ture medium will be substantially covered with a rowth of white mycelial fungi of a white silky luster. If the process is stopped at this point and the pro uct dried as before described, it is the new ko'i.

If the process be not stoppe as last described, a further growth will gradually assume, first, a ellowish and subsequently a greenish ye ow appearance. After a lapse of 64: to 74 hours, more or less, from the beginning, this growth will -have attained its maximum. If the product be dried at this stage, it is the new moyashi.

The incubator is so arranged as to saturate the air with aqueous vapor to a maximum so as to keep the culture always moist; also, it should be well ventilated so as to permit the escape of carbon dioxid and supalso, as large a surace of the culture medium as possible should be exposed to the air. This can be done by spreading on trays to a thickness of about one to three inches. Within 12 to 16- hours, the growth starts and the temperature goes gradually up until it not infre uently reaches 42 to 43 C. It is desira le to keep the temperature as near as possible to 30 C. since too high a temperature will, for a while, slacken the growth of the fungus; but more or less igher temperature than 30 C. does not considerably affect the final result except- 111% as it entails a loss of time.

be reason for the conditions last described is, because Aspergillus oryzw fungi need a sufiicient sup ly of oxygen and a large culture surface d urin rowth. These conditions, however, woul Iced, as an unavoidable consequence, to the mass being infected and contaminated with foreign fungi and bacteria were their growth not temporarily or permanently prevented by the presence of that proportion of antiseptic to which the Aspergillus org 2w fungi have been rendered immune by the acclimatization.

Attempts heretofore made to prevent the growth of such foreign fungi, such as the filtering of the air entering the incubator, have failed. This failure is in part due to the practical necessity of opening the chamber and also of use of utensils which it is impossible to keep clean.

As some of the antiseptics which may be substituted for formaldehyde, I may mention h drofluoric, benzoic or salicylic acid. By this acclimatization the spores may be made immune to hydrofluoric acid in the proportion of 1 to 2000 by weight of the moist culture medium and benzoic or salicylic acid in the proportion of 1 to 500.

I prefer formaldehyde because itis dissipated from the mass by the time the koji growth is complete, by partial volatilization and also because of its combining power with protein matters in which combinations it has lost its antiseptic property.

In place of the wheat bran, I may substitute comminuted grains of cereals but the bran of cereals I prefer on account of its volume which afiords larger surface for the growth of the fungi. I do not, however, limit myself to these substances since I believe that any substance which at the same time presents suflicient surface and affords sufficient nutriment'for the fungi me be employed as the culture medium, suc as dried distillers or brewers slop etc.

I believe that I am the first one to discover or produce a fungus for enzyms having starch converting (i. e. convertin starch into su ar) power and at the same time substantia ly immune from the action of a proportion of antiseptic incom atible with the owth of other f lieve that I am t e first one to acclior acteria. I also matize any substance having starch convertinglpower against antiseptic influence.

- y new product in any of its forms may be recognized by possession of the following characteristics in combination, namely: It possesses diastatic power and when tested in comparison with ordinary fungus of the same species it displays an abnormal resistance against the influence of an antiseptic, such as formaldehyde.

My new culture of diastatic fungus has the strength of converting from ten to twenty times its weight of starch; the wei ht of the culture being taken dry.

aving thus described my invention, I claim as new:

1. The improvement in the manufacture of diastase producing substances which ,con-

sists in growing repeated generations offungi possessing enzyms having starch converting power in the presence of increas-.

ing proportions of an antiseptic.

2. The improvements in the manufacture of diastase producing substances which consists in growing repeated generations of Aspergillus in the presence of increasing proportions of an antiseptic.

3. The improvement in the manufacture of diastas'e producing substances which con- 1 portions of formaldehyde.

5. The improvement in the manufacture of moyashi which consists in growing acclimatized Aspergillus fungi in the presence of an antiseptic.

6. As an article of manufacture, a moya shi characterized by the possession of power of producing fungus possessing enzyms having diastatic power combined with an abnormal resistance to antiseptic power.

In testimony whereof, I have hereunto signed my name in the presence of two subscribing witnesses.

J OKICHI TAKAMIN E.

Witnesses:

M. E. MON INCH, ARTHUR GOAT.

Correctlons in Letters Patent No.1,148,938.

It is hereby certified that in Letters Patent No. 1,148,938, granted August 3, '1915, upon the application of Jokichi Takamine, of New York, N. Y., for an improvement in Diastetic Products and the Processes for Producing the Same, errors appear in the printed specification requiring correctiop as follows: Page 2, line 29, after the date, strike out the comma and insert a period and the words It can also be used, thus making a new sentence; page 3, line 23, claim 2, for the word improvements read improvement; and that the said Letters Patent should he read with these corrections therein that the same may conform to the record of the case in the Patent Ofiioe.

Signed and sealed this 31st day of August, A. D., 1915.

time J T. NEWTON,

Acting Commissioner of Patchts. 

